Objective To study the effect of Iguratimod on the collagen secretion of 3T6 cells. Method The median toxic concen- tration of Iguratimod was determination by MTT Assay. Iguratimod' s median toxic concentration to 3T6 cell was about 1142.6mg/L. Taking into the consideration of the cells performance after administration under spectacles, we chosen the administration concentration as 334.8 mg/L(DMS0:1?l/256?l) ,167.4mg/L,83.7mg/L and 41.85 mg/L. After administration of 12 hours, 24 hours, 36 hours and 48 hours, the supernatant was collected for assayed respectively. The collagens content was detected by the method of chloramine T. Results The collagen concentration of the drug concentration 334.375mg/L group increased (P

Objective To investigate the expressions and significance of interleukin 8 (IL 8) in patients with ankylosing spondylitis (AS). Methods Using microarray and RT PCR to detect the expressions of IL 8 in peripheral blood mononuclear cells (PBMC), synovial fluid mononuclear cells (SFMC) and synovial cells in patients with active AS. Comparing the results with those in PBMC of both healthy volunteers and patients with rheumatoid arthritis (RA), and also with those in synovial cells obtained from patients undergoing knee operations for trauma. Results The expressions of IL 8 in PBMC and SFMC of patients with active AS were significantly higher than those in PBMC of the healthy controls. The expression of IL 8 in PBMC of RA patients was also higher than that of healthy volunteers. Higher expression of IL 8 in the synovial cells were detected in AS than that of patients with knee joint trauma. Conclusions The expression of IL 8 is high in patients with AS. The result may indicate that IL 8 is possibly one of the important mediators of inflammation in AS, and it may mediate the induction and development of inflammation in synovium or other tissues involved in AS

Objective To consecutively understand the national clinical testing status and to reassure the quality-control of autoantibody detection. Methods Letter or telephone notification were conducted to the participating hospitals or departments. Autoantibodies for quality control survey included anti-nuclear anti-body (ANA), anti-double-stranded DNA (anti-dsDNA), anti-extractable nuclear antigens (A-ENA), anti-mitochondria antibody (AMA)/anti-smooth muscle antibody (ASMA), and anti-citrulline antibody (anti-CCP). Each had 3 control samples, and altogether 15 samples for testing. Sample distribution and data analysis were double-blinded. Results One hundred and two hospitals/departments participated in the national quality-control survey. The accurate rate for this survey was 70%, 88%, 93%, 85%, 79% respectively for ANA, anti-dsDNA, AMA, ASMA and anti-CCP. Anti-ENAs were further divided into anti-RNP, Sm, SSA, SSB and Scl-70 subgroups, and the accurate rate was 88%, 77%, 92%, 93% and 87% respectively. Conclu-sion Compared to the previous 4 national surveys, the accurate rates of ANA, anti-dsDNA, anti-ENAs, AMA in our country's autoantibody testing is improved, and the detection rate of ASMA and anti-CCP antibody is also increased. More hospitals and testing items can test these autoantibodies, which implies that autoantibody testing status has been improving in our country.

Objective Previous study had shown that signal transduction was changed when Hela cells were transfected with HLA-B27gene.An increase of monocyte chemotactic protein-1(MCP-1)of Hela cells was observed.To determine the role of MCP in the pathogenesis of ankylosing spondylitis(AS),expression of MCP-1,2,3and4in peripheral blood mononuclear cells(PBMC),synovial fluid mononuclear cells(SFMC)and synovial tis-sues of AS patients were tested.Methods Gene expression profiles of PBMC from AS patients and healthy volun-teers were determined by cDNA microarray with1176target gene filter.The differentiated expressed gene MCP-1in PBMC,SFMC and synovial tissue of AS patients were confirmed by semi-quantitative RT-PCR.Results The gene expression profile of SFMC of AS patients was significantly different from those of PBMC from AS and PBMC from healthy volunteers.The MCP-1level was positively correlated with MCP-3(r=0.76,P=0.03).The expressions of MCP-1were higher in synovial tissues of AS than those of healthy volunteers(P=0.0035).MCP-1levels in monocytes of AS patients and control subjects were increased after LPS stimulation for4hours.Conclusions There is increased expression of MCP-1in SFMC and synovial tissue of AS patients.The results indicate that MCP-1may play a potential key role in the homing of cells migrating from blood to joint and in the pathogenesis of joint inflammation in AS patients.

BACKGROUND: Human leukocyte antigen (HLA)-B27 is closely connected to the occurrence of some rheumatic diseases such as ankylosing spondylitis and can be used as an important factor for evaluating the diagnosis of ankylosing spondylitis. Immunomagnetic separation and enzymelinked immunosorbent assay (IMS-ELISA) has been applied to the detection of HLA-B27.OBJECTIVE: To explore the accuracy, sensitivity and specificity of IMSELISA for detecting HLA-B27 and its value in the auxiliary diagnosis of ankylosing spondylitis.DESIGN: A clinical trial in comparison with the gold standard.SETTING: Departments of Rheumatology and Clinical Laboratory, Third Affiliated Hospital of Sun Yat-sen University.PARTICIPANTS: Eighty-six patients suffering from low back pain and/or arthritis who were treated for the first time in Department of Rheumatology from December 2002 to April 2003. Inclusion criteria: ① Presence of manifestations of low back pain and/or arthritis; ② Thorough documentation of clinical and other examinations; ③ Informed consent to HLA-B27 examination; ④ Treatment for the first time in the Third Affiliated Hospital of Sun Yet-sen University. Those with other serious diseases or with incomplete record of clinical and/or accessory examinations were excluded. The 86 patients included 56 male and 30 female patients aged from 12 to 65 years.METHODS: Blood sample was detected for HLA-B27 by both IMS-ELISA and microlymphocytotoxicity test, and the latter was selected as the gold standard. The coincidence rate of the results detected by the two methods as well as the sensitivity, specificity, positive and negative predictive values of IMS-ELISA were calculated.MAIN OUTCOME MEASURES: ① The coincidence rate of the results of the two methods. ② The sensitivity and specificity of IMS-ELISA for detecting HLA-B27.RESULTS: None of the patients was lost. For the 33 patients with ankylosing spondylitis, the positivity rate of IMS-ELISA (90.9％) was higher than that of microlymphocytotoxicity test (87.9％), but the difference was not statistically significant (P＞0.05). The total coincidence rate of the two methods was 93.0％ in all the 86 patients. The sensitivity, specificity, positive and negative predictive values of IMS-ELISA were 90.0％, 95.7％,94.7％ and 91.7％ respectively.CONCLUSION: Both IMS-ELISA and microlymphocytotoxicity test are capable of reliable examination of HLA-B27 with high sensitivity and specificity.

BACKGROUND: Spondyloarthropathies(SpA) are a group of interrelated disorders that involve the spine, peripheral joints, and tissues around or outside the joints. SpA are most common in young males, and both the incidence and the mutilation rate are very high. It is important to evaluate the disease activity of patients with SpA since the evaluation can help in therapeutic selection and prognostic prediction of SpA. But up to now there still has not been a widely clinically acceptable criterion to evaluate the disease activity of SpA.OBJECTIVE: To study the worth of color Doppler(CD) in assessing the disease activity of patients with SpA.DESIGN: Non-random and self-control clinical study based on the patients.SETTING: All patients recruited in this study were from department of rheumatology of the Third Affiliated Hospital of Sun Yat-sen University. And all the ultrasonic examinations were conducted in the department of ultrasound of the same hospital.PARTICIPANTS: Thirty(24 males and 6 females) patients with SpA who met the European SpA Study Group criteria for the classification of SpA issued in 1991 and consented to be examined by CD were included.METHODS: CD was used to determine the blood perfusion at 18 major entheses of lower limbs of each patient with a semi-quantitative method. Blood sample was also taken from the patients for the examination of erythrocyte sedimentation rate(ESR) and C reactive protein(CRP) . The disease activity of the patients was also evaluated with the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI).MAIN OUTCOME MEASURES: To observe the inter-correlations between entheseal blood perfusion, ESR, CRP and BASDAI.RESULTS: 80% of the 30 patients were detected to have positive perfusion in at least one enthesis of the 18 examined entheses, and all together 84 entheses were found to have positive perfusion among the 540 entheses being detected(positive rate 15.6% ) . The sum of entheseal perfusion detected in all the 18 sites was statistically correlated with the sum of patients' ESR,CRP, BASDAI respectively with the correlation coefficients of 0. 658, 0. 428and 0. 821 (all P ＜ 0.05) . ESR and CRP were also statistically correlated with BASDAI and their correlation coefficients were 0. 675 and 0. 368(Both P ＜ 0.05) . The sum of perfusion detected in the 18 entheses sites was correlated with BASDAI better than ESR or CRP was.Detecting the.entheseal perfusion by CD can help to determine the disease activity of patients with SpA, and then to guide selecting therapeutic strategy. It is also important for following up and prognosis.

BACKGROUND: Seronegative spondyloarthropathies(SpA) are a group of diseases involving the spine, peripheral joints and structures around the joints that mostly occur in adult males with high morbidity and disability rate. Presently, it is widely believed that enthesis is the initiated site of SpA so that enthesitis is an important pathologic base and one of the characteristic clinical manifestations of SpA, however, there is no specific clinical manifestation enthesitis.OBJECTIVE: To compare the difference on entheseal blood flow between health volunteers and patients with SpA and explore the value of color Doppler(CD) in assessing the pathological change of enthesis.DESIGN: Non-randomized case controlled study to patients SETTING: Department of Rheumatology, Department of Ultrasound of a university, Department of Rheumatology of General Hospital of Chinese PLA.PARTICIPANTS: Totally 30 patients with SpA admitted into Third Affilated Hospital of Sun Yat-sen University during August 2001 to April 2002 and willing to take CD examination were selected as case group. All of them met the classification criteria of SpA set up by the European SpA Study Group in 1991 with 24 males and 6 females. Thirty healthy workers and students working or studying in the same Hospital were enrolled as volunteers for the control group, none of them had any histories of arthropathy or chronic lower back pain, and there were 22 males and 8 females among them.INTERVENTIONS: Color Doppler was used to examine the blood flow of 18entheses in lower limbs of every healthy volunteers and SpA patients and semi-quantitative method was used to classify the blood flow.MAIN OUTCOME MEASURES: Blood flow of enthesesRESULTS: Among 30 healthy volunteers, 5 cases(16.7% ) had been detected blood flow from at least one enthesis. From the 540 entheses being examined, 9entheses were detected blood flow signals of grade Ⅰ, the positive rate was 1.7%. While among 30 SpA patients, 24(80.0%) had at least one enthesis being detected for positive blood flow. Eighty-four of totally 540 exanined entheses were found for positive blood flow signals with a positive rate of 15.6%, and the number of entheses with grade Ⅰ, Ⅱ and Ⅲ of blood flow were 19, 33 and 32 respectively among the 84 positive entheses. There was significantly different in the positive rate of blood flow and classification of entheses between SpA patients and healthy volunteers by x2 test (P＜ 0.05).CONCLUSION: Colour Doppler examination is an effective way to detect the blood flow of enthesis. There were blood flow abnormity in the entheses of lower limbs in patients with SpA.

Objective To search for the genetic and molecular immunity basis of CXCR-1 associated pathogenesis in ankylosing spondylitis (AS) patients. Methods Sequencing analysis was used to detect mutation in the exonic, junctional and promoter sequences of CXCR-1 which might be related with ankylosing spondylitis; the hydrophobicity, conservation and evolutionary distance of the mutated amino acids were also analyzed. Results Six affected individuals in the family were detected with a novel mutation Arg192Gly. The glycine at 192 codon was highly conserved in different species. Arginine and glycine had quite distinct hydrophobicity and BLOSUM score. Conclusion The mutation CXCR-1 (Arg192Gly) detected in these patients might be involved in genetic and molecular immunity mechnisms of ankylosing spondylitis.

Objective To investigate the reasons for delayed diagnosis of ankylosing spondylitis(AS) in a Chinese population.Methods Three hundred and eight patients fulfilled the 1984 Modified New York criteria of AS were enrolled.They were interviewed in person or by telephone by rheumatologists for 13 ques-tions.Results Of the 308 AS patients.238(75%)completed 13 questions.Among these 238 patients,male to female ratio was 6 to 1.The average age at AS onset was 22.1 years.Those aged under 15 years at disease onset Was 18.1%.and between 15 and 39 years was 79.0%while over 39 years was 2.9%.Of these 238 pa-tients.27(23.9%)had family history of AS.84.9% of the patients were HLA-B27 positive.The average dura-tion of delayed diagnosis in HLA-B27(+)and HLA-B27(-)were 70.1 and 88.1 months respectively,which was not significant statistically.Among those delayed over 10 vears,24 AS patients were HLA-B27(+)and 10 were HLA-B27(-),which was statistically significant(P=0.012).66.4% of 238 patients were misdiagnosed, of which 23.4%were diagnosed as arthritis associated with rheumatic fever,22.9% as fatigue and 20.3%as inter-vertebrate disc.45.0%were misdiagnosed by one physician while 2 1.4%were misdiagnosed for several times,and the average length of delay diagnosis was 72±68 months.Seven cases,although the diagnosisi was being delaved for more than 10 years the initial diagnosis was correct. Conclusion Delayed diagnosis of AS is common in China.The major reasons for delayed diagnosis are HLA-B27 negative and lack of X-ray changes of sacroiliac joint at the initial visit.

Objective To study T lymphocyte subsets and expression of costimulatory molecule CD154 on T-cells in peripheral blood from patients with ankylosing spondylitis and their changes after treated with Enbrel. Methods Sixty-six patients with AS(39 active and 27 inactive, 35 axial and peripheral joint involvement and 31 axial involvement only), 30 patients with rheumatoid arthritis(RA), 30 healthy volunteers were analyzed. The expression of CD154 on CD3+ T cell as well as T-cells subsets were evaluated using flow cytometry respectively. The changes of the expression of costimulatory molecule CD154 in 39 active AS patients(Enbrel treatment or placebo treatment) were observed in a randomized, double-blind, placebothan that of healthy volunteers, and CD154 expression on CD3+ T cells in peripheral blood in AS patients was on CD3+ T cells in the peripheral blood of active AS or AS patients with peripheral joint involvement were significantly higher than those in inactive or axial involvement only AS patients(P＜0.05), and CD154 on CD3+ placebo, in AS patients group, there was significant reduction in CD154 expression on CD3+ T cells in Enbrel group at week 6(P＜0.05), there was no significant difference between Enbrel group and healthy volunteers at week 6(P＞0.05). Conclusion T lymphocyte subsets are significantly abnormal and CD154 is overexpressed on T-cells in peripheral blood of patients with AS. A six-week course of treatment with Enbrel in active AS can induce a down-regulation of the expression of CD154 on T cells.